Detection of antibiotic-related genes from bacterial biocontrol agents with polymerase chain reaction

2006 ◽  
Vol 52 (5) ◽  
pp. 476-481 ◽  
Author(s):  
Y Zhang ◽  
W G.D Fernando ◽  
T R. de Kievit ◽  
C Berry ◽  
F Daayf ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Carboxylic Acid ◽  
Bacillus Thuringiensis ◽  
Resistance Gene ◽  
Field Experiments ◽  
Significant Inhibition ◽  
Chain Reaction ◽  
Pseudomonas Spp ◽  
Zwittermicin A ◽  
Polymerase Chain

Pseudomonas chlororaphis PA23, Pseudomonas spp. strain DF41, and Bacillus amyloliquefaciens BS6 consistently inhibit infection of canola petals by Sclerotinia sclerotiorum in both greenhouse and field experiments. Bacillus thuringiensis BS8, Bacillus cereus L, and Bacillus mycoides S have shown significant inhibition against S. sclerotiorum on plate assays. The presence of antibiotic biosynthetic or self-resistance genes in these strains was investigated with polymerase chain reaction and, in one case, Southern blotting. Thirty primers were used to amplify (i) antibiotic biosythetic genes encoding phenazine-1-carboxylic acid, 2,4-diacetylphloroglucinol, pyoluteorin, and pyrrolnitrin, and (ii) the zwittermicin A self-resistance gene. Our findings revealed that the fungal antagonist P. chlororaphis PA23 contains biosynthetic genes for phenazine-1-carboxylic acid and pyrrolnitrin. Moreover, production of these compounds was confirmed by high performance liquid chromatography. Pseudomonas spp. DF41 and B. amyloliquefaciens BS6 do not appear to harbour genes for any of the antibiotics tested. Bacillus thuringiensis BS8, B. cereus L, and B. mycoides S contain the zwittermicin A self-resistance gene. This is the first report of zmaR in B. mycoides.Key words: Pseudomonas, Bacillus, biocontrol, antibiotic genes.

Characterization ofBacillus thuringiensismutants and natural isolates by molecular methods

1997 ◽  
Vol 43 (5) ◽  
pp. 403-410 ◽  
Author(s):  
Yong Chul Jung ◽  
Sung Uk Kim ◽  
Song Hae Bok ◽  
Ho Yong Park ◽  
Jean-Charles Côté ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacillus Thuringiensis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Rice Grain ◽  
Chain Reaction ◽  
Hyphantria Cunea ◽  
Natural Isolates ◽  
Grain Dust ◽  
Polymerase Chain

Two Bacillus thuringiensis var. kurstaki HD-1 mutants, two Bacillus thuringiensis var. israelensis HD-500 mutants, and four rice grain dust isolates were characterized using microscopic examination and protein profiles of purified crystals on sodium dodecyl sulfate – polyacrylamide gel electrophoresis. Specific detection of cryI- and cryIV-type genes was performed in a polymerase chain reaction using cryI and cryIV-specific oligonucleotide primers. The cry-type genes under study consisted of cryIA(a), cryI(A)b, cryI(A)c, cryIB, and cryIV. Presence or absence of the cryI- and cryIV-type genes was further confirmed by Southern blotting followed by hybridization with specific cryI and cryIV gene fragments. A genetically modified strain of B. thuringiensis var. kurstaki HD-1, called OZK-13 and obtained following mutagenesis with ozone, was shown to contain cryIA(a), cryIA(b), and cryIA(c) genes. Another kurstaki HD-1 mutant, called NGK-13 and obtained following treatment with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), was shown to have lost the cryIA(b) gene while retaining the cryIA(a) and cryIA(c) genes. NGI-23-1, an oligosporogenous–multicrystalliferous mutant of B. thuringiensis var. israelensis (Bti) HD-500, obtained following treatment with MNNG contained cryIV-type genes. NGI-22, an oligosporogenous–acrystalliferous mutant of Bti HD-500, contained no cryI- nor cryIV-type genes. The rice grain dust isolate BT-285 contained the cryIA(a) and cryIA(c) genes. Isolate BT-14 contained only the cryIA(c) gene, whereas isolate BT-209 contained cryIA(a), cryIA(b), and cryIB genes. Isolate BT-205 contained no cryI- nor cryIV-type genes. Bacillus thuringiensis mutants and natural isolates shown to contain cryI-type genes were tested for their insecticidal activities in a series of bioassays against Hyphantria cunea Drury (Lepidoptera: Arctiidae). All cryI-carrying strains were toxic against the insect larvae. BT-205 was also tested and exhibited no toxicity against the insect larvae.Key words: Bacillus thuringiensis, δ-endotoxin crystal, cry-type genes, polymerase chain reaction.

Identification of an RAPD marker for the crown rust resistance gene Pc68 in oats

Genome ◽  
1993 ◽  
Vol 36 (5) ◽  
pp. 818-820 ◽  
Author(s):  
G. A. Penner ◽  
J. Chong ◽  
C. P. Wight ◽  
S. J. Molnar ◽  
G. Fedak
Keyword(s):  
Polymerase Chain Reaction ◽  
Resistance Gene ◽  
Genomic Dna ◽  
Bulk Segregant Analysis ◽  
Rust Resistance ◽  
Crown Rust ◽  
Rust Resistance Gene ◽  
Chain Reaction ◽  
Polymerase Chain Reaction Technology ◽  
Polymerase Chain

The feasibility of using bulk segregant analysis to identify molecular markers for disease resistance genes in oats was investigated, utilizing random primers in conjunction with polymerase chain reaction technology. Random primers were screened for the amplification of polymorphic DNA fragments on two pools of genomic DNA isolated from plants that were homozygous for the presence and absence of the crown rust resistance gene Pc68. Ten primers were identified that amplified polymorphic DNA fragments. Of these, one was tightly linked, in repulsion, to the target gene, while the other nine were not linked to this trait. The relatively low cost of polymerase chain reaction technology, coupled with rapid leaf disc genomic DNA extraction techniques should result in the effective use of this linked marker in oat breeding selection programs.Key words: RAPD, molecular marker, oats Puccinia coronata, bulk segregant analysis, Pc68.

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